NADPH oxidase activation contributes to native low-density lipoprotein-induced proliferation of human aortic smooth muscle cells.
NADPH oxidase activation contributes to native low-density lipoprotein-induced proliferation of human aortic smooth muscle cells.
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DOI：
10.1038/emm.2015.30
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发表时间：
2015-06-12
影响因子：
12.8
通讯作者：
Ryoo, Sungwoo
中科院分区：
文献类型：
--
作者：
Park, Il Hwan;Hwang, Hye Mi;Jeon, Byeong Hwa;Kwon, Hyung-Joo;Hoe, Kwang Lae;Kim, Young Myeong;Ryoo, Sungwoo
天然低密度脂蛋白（nLDL）血浆浓度升高与血管平滑肌细胞（VSMC）活化和心血管疾病相关。我们研究了超氧化物生成的机制及其对nLDL刺激引起的病理生理细胞增殖的贡献。使用光泽精诱导的化学发光来测量nLDL诱导的人主动脉平滑肌细胞（hAoSMC）中的超氧化物产生。在nLDL刺激的hAoSMC和hAoSMC匀浆以及制备的膜组分中，烟酰胺腺嘌呤二核苷酸磷酸（NADPH）可增加超氧化物的产生，NADPH氧化酶抑制剂可降低超氧化物的产生。细胞外信号调节激酶1/2（Erk 1/2）、蛋白激酶C-θ（PKCθ）和蛋白激酶C-β（PKCβ）在nLDL刺激3 min内磷酸化并最大程度激活。磷酸化Erk 1/2丝裂原活化蛋白激酶、PKCθ和PKCβ刺激p47 phox和p22 phox之间的相互作用;这些相互作用被MEK和PKC抑制剂（分别为PD 98059和calphostin C）阻止。这些抑制剂降低nLDL依赖性超氧化物的产生，并阻止p47 phox易位到膜上，如荧光成像和细胞分级分离实验所示。增殖试验表明，针对p47 phox的小干扰RNA以及超氧化物清除剂和NADPH氧化酶抑制剂阻断nLDL诱导的hAoSMC增殖。nLDL刺激C57 BL/6 J小鼠去内皮化主动脉环增加二氢乙啶荧光并诱导p47 phox易位，其被PD 98059或calphostin C阻断。从p47 phox −/−小鼠分离的主动脉SMC（mAoSMC）对nLDL刺激没有反应。此外，NADPH氧化酶1（Nox 1）负责nLDL刺激的hAoSMC中超氧化物的产生和细胞增殖。这些数据表明，NADPH氧化酶活化有助于nLDL刺激的hAoSMC中的细胞增殖。
Elevated plasma concentration of native low-density lipoprotein (nLDL) is associated with vascular smooth muscle cell (VSMC) activation and cardiovascular disease. We investigated the mechanisms of superoxide generation and its contribution to pathophysiological cell proliferation in response to nLDL stimulation. Lucigenin-induced chemiluminescence was used to measure nLDL-induced superoxide production in human aortic smooth muscle cells (hAoSMCs). Superoxide production was increased by nicotinamide adenine dinucleotide phosphate (NADPH) and decreased by NADPH oxidase inhibitors in nLDL-stimulated hAoSMC and hAoSMC homogenates, as well as in prepared membrane fractions. Extracellular signal-regulated kinase 1/2 (Erk1/2), protein kinase C-θ (PKCθ) and protein kinase C-β (PKCβ) were phosphorylated and maximally activated within 3 min of nLDL stimulation. Phosphorylated Erk1/2 mitogen-activated protein kinase, PKCθ and PKCβ stimulated interactions between p47phox and p22phox; these interactions were prevented by MEK and PKC inhibitors (PD98059 and calphostin C, respectively). These inhibitors decreased nLDL-dependent superoxide production and blocked translocation of p47phox to the membrane, as shown by epifluorescence imaging and cellular fractionation experiments. Proliferation assays showed that a small interfering RNA against p47phox, as well as superoxide scavenger and NADPH oxidase inhibitors, blocked nLDL-induced hAoSMC proliferation. The nLDL stimulation in deendothelialized aortic rings from C57BL/6J mice increased dihydroethidine fluorescence and induced p47phox translocation that was blocked by PD98059 or calphostin C. Isolated aortic SMCs from p47phox−/− mice (mAoSMCs) did not respond to nLDL stimulation. Furthermore, NADPH oxidase 1 (Nox1) was responsible for superoxide generation and cell proliferation in nLDL-stimulated hAoSMCs. These data demonstrated that NADPH oxidase activation contributed to cell proliferation in nLDL-stimulated hAoSMCs.
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影响因子：
4.8
作者：
Mahadev, K;Zilbering, A;Goldstein, BJ
通讯作者：
Goldstein, BJ
影响因子：
8.3
作者：
SACHINIDIS, A;MENGDEN, T;VETTER, W
通讯作者：
VETTER, W
影响因子：
4.8
作者：
Ushio-Fukai, M;Alexander, RW;Griendling, KK
通讯作者：
Griendling, KK
影响因子：
15.9
作者：
Barry-Lane, PA;Patterson, C;Runge, MS
通讯作者：
Runge, MS
影响因子：
20.1
作者：
Seshiah, PN;Weber, DS;Griendling, KK
通讯作者：
Griendling, KK